Three-dimensional electron microscopy of the reverse gyrase from Sulfolobus tokodaii |
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Authors: | Matoba Kyoko Mayanagi Kouta Nakasu Syo Kikuchi Akihiko Morikawa Kosuke |
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Affiliation: | Department of Structural Biology, Biomolecular Engineering Research Institute (BERI), 6-2-3 Furuedai, Suita-city, Osaka 565-0874, Japan. |
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Abstract: | Reverse gyrase is a type IA topoisomerase, found in various hyperthermophiles and promotes ATP-dependent positive supercoiling of DNA. Electron microscopy combined with single particle analyses revealed the three-dimensional structure of the DNA-free Sulfolobus tokodaii reverse gyrase and two-dimensional average images of both the protein alone and that complexed with double-stranded DNA. The 23A resolution map exhibited a parallelogrammatic morphology of 110 x 87 x 43A, which is in good agreement with the crystal structure of the Archaeoglobus fulgidus reverse gyrase. The average image of the complex revealed that the monomeric enzyme binds DNA duplex. Together with this average image of the complex, the three-dimensional map implies that, at the beginning of the supercoiling reaction, DNA is bound within a 10-20A wide cleft in the helicase-like domain. We also speculate that DNA may pass through a 20A wide hole at the end of the cleft. |
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Keywords: | Archaea DNA topology Electron microscopy Positive supercoiling Reverse gyrase Topoisomerase Single particle analysis Sulfolobus tokodaii Thermophile Three-dimensional structure |
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